Ectopic hCG-beta secreted by DoT and CaSki human carcinoma cell lines contains two components, ectopic hCG-beta-I and ectopic hCG-beta-II. Ectopic hCG-beta-II lacks the C-terminal peptide (CTP), based on lack of recognition by antibodies against CTP, lack of susceptibility to thermolysin (which normally cleaves the CTP), and lack of binding of the desialylated derivative to Arachis hypogaea (AH) lectin (specific for Gal-GalNAc of the O-linked oligosaccharides on the CTP). Our studies will test the hypothesis that there are at least two hCB-beta peptides synthesized ectopically by cervical carcinoma. Serum and urine of cervical carcinoma patients will be analyzed for differential recognition by antisera to either the core portion of CTP or hCG-beta. Pulse-chase experiments will be performed with both DoT and CaSki cell lines to determine if a precursor-product relationship exists between ectopic hCG-beta-II. The isolation procedure will rely on purification by anti-hCG-beta affinity chromatography followed by immune precipitation with antiserum to CTP and/or AH chromatography of desialylated ectopic hCG-beta (AH binds desialylated ectopic hCG-beta-I but not desialylated ectopic hCG-beta-II), and HPLC. Corroborative evidence for lack of CTP (which is rich in Ser and Pro but lacks Met) on ectopic hCG-beta-II will be obtained from amino acid composition and from ratio of 3H-Pro/35S-Met, before and after thermolysin digestion of ectopic hCG-beta-I and ectopic hCG-beta-II biosynthesized by DoT and CaSki cells.